Temporal ecological assessment of oil contaminated soils before and after bioremediation

Ecotoxicity methods were used to assess different soil and oil combinations before, during and after laboratory bioremediation with associated hydrocarbon analysis. Heavy, medium and light crude oil (API gravity 14, 30, and 55) was spiked (ca. 5% w/w) into two sandy soils in the laboratory having organic carbon concentrations of 0.3 (Norwood) and 4.7% (Norwood/Baccto). The earthworm (Eisenia fetida) 14-d lethality assay, the modified Microbics Microtox Solid-Phase assay, and the 14-d plant seed germination and growth assays using corn, wheat and oats, were spiked and tested during a 360-d laboratory remediation. Eisenia was the most sensitive of the three methods utilized with survival increasing throughout bioremediation with fastest toxicity reduction in the high carbon Norwood/Baccto soils where LC50's were 100% or greater at the end of 90-d whereas, > 150-d were required to achieve a similar result in the low carbon soil. Analysis of the undiluted treatments with oily soil alone showed that earthworm survival was high after 90-d in all high organic carbon soils, and after eight months in the low carbon soils, except for the Norwood soil-light oil treatment, which required 360-d to achieve 100% survival. The Microtox assay was less sensitive with EC50's 100% or greater observed after 90-d in high carbon soils and after 240-d for all low carbon soils. After bioremediation, no effects on seed germination were observed, although some plant growth inhibition effects remained. There was no direct correlation between total petroleum hydrocarbon concentrations and toxicity.     

中原油田石油污染土壤原位生物修复技术实验研究

通过实验室选择性富集培养,从中原油田石油污染土壤中获得了能以中原原油为碳源快速生长的石油降解菌群。结合黑麦草（Ryegrass）和苜蓿（Alfalfa）,采用该降解菌群对原油污染土壤进行了原位生物联合修复实验。接入降解菌的实验区分种植黑麦草、种植苜蓿、未种植区,另设黑麦草区和空白区。经过99 d的生物修复,石油烃累计降...     

Ecorisk evaluation and treatability potential of soils contaminated with petroleum hydrocarbon-based fuels

We used a series of toxicity tests to monitor oil degradation in the Kuwaiti oil lakes. Three soils from different locations with a history of hydrocarbon contamination were treated in bench-scale microcosms with controlled nutrient amendments, moisture content, and temperature that had promoted mineralization of total hydrocarbon and oil and grease in a preliminary study. Two hundred days of bioremediation treatment lowered hydrocarbon concentration to below 2 and 5 mg g(-1) for soils A and B, respectively, while in soil C hydrocarbon concentration remained at 12 mg g(-1). Although 85% of the total petroleum hydrocarbons (TPHs) in soil A were reduced 50d after treatment, results of the seed germination and Microtox tests suggested an initial increase in toxicity, indicating that toxic intermediary metabolites may have formed during biodegradation. Also, the significant decrease of TPHs and corresponding high toxicity levels were noted in soil B 200d after bioremediation. Clearly, toxicity values, and not just hydrocarbon concentration, are a key factor in assessing the effectiveness of bioremediation techniques. Field chemistry data showed a significant reduction in hydrocarbon levels after the biological treatment. We concluded that the toxicity assessment of the contaminated soil with a battery of toxicity bioassays could provide meaningful information regarding a characterization procedure in ecological risk assessment.     

Ex situ bioremediation of a soil contaminated by mazut (heavy residual fuel oil)--a field experiment

Mazut (heavy residual fuel oil)-polluted soil was exposed to bioremediation in an ex situ field-scale (600 m(3)) study. Re-inoculation was performed periodically with biomasses of microbial consortia isolated from the mazut-contaminated soil. Biostimulation was conducted by adding nutritional elements (N, P and K). The biopile (depth 0.4m) was comprised of mechanically mixed polluted soil with softwood sawdust and crude river sand. Aeration was improved by systematic mixing. The biopile was protected from direct external influences by a polyethylene cover. Part (10 m(3)) of the material prepared for bioremediation was set aside uninoculated, and maintained as an untreated control pile (CP). Biostimulation and re-inoculation with zymogenous microorganisms increased the number of hydrocarbon degraders after 50 d by more than 20 times in the treated soil. During the 5 months, the total petroleum hydrocarbon (TPH) content of the contaminated soil was reduced to 6% of the initial value, from 5.2 to 0.3 g kg(-1) dry matter, while TPH reduced to only 90% of the initial value in the CP. After 150 d there were 96%, 97% and 83% reductions for the aliphatic, aromatic, and nitrogen-sulphur-oxygen and asphaltene fractions, respectively. The isoprenoids, pristane and phytane, were more than 55% biodegraded, which indicated that they are not suitable biomarkers for following bioremediation. According to the available data, this is the first field-scale study of the bioremediation of mazut and mazut sediment-polluted soil, and the efficiency achieved was far above that described in the literature to date for heavy fuel oil.     

Biodegradation of endosulfan by Mortieralla sp. strain W8 in soil: Influence of different substrates on biodegradation

To examine the bioremediation potential of Mortierella sp. strain W8 in endosulfan contaminated soil, the fungus was inoculated into sterilized and unsterilized soil spiked with endosulfan. Wheat bran and cane molasses were used as substrates to understand the influence of different organic materials on the degradation of endosulfan in soil. Strain W8 degraded α- and β-endosulfan in both sterilized and unsterilized soil. In unsterilized soil with wheat bran+W8, α- and β- endosulfan were degraded by approximately 80% and 50%, respectively after 28 d incubation against the initial endosulfan concentration (3 mg kg(-1) dw). The corresponding values for α- and β-endosulfan degradation with wheat bran only were 50% and 3%. Endosulfan diol metabolite was detected after 14 d incubation in wheat bran+W8 whereas it was not found with wheat bran only. Production of endosulfan sulfate, the main metabolite of endosulfan, was suppressed with wheat bran+W8 treatment compared with wheat bran only. It was demonstrated that wheat bran is a more suitable substrate for strain W8 than cane molasses. Wheat bran+W8 is a superior fungus and substrate mix for bioremediation in soil contaminated with endosulfan.     

The application of bioassays as indicators of petroleum-contaminated soil remediation

Bioremediation has proven successful in numerous applications to petroleum contaminated soils. However, questions remain as to the efficiency of bioremediation in lowering long-term soil toxicity. In the present study, the bioassays Spirotox, Microtox, Ostracodtoxkit F, umu-test with S-9 activation, and plant assays were applied, and compared to evaluate bioremediation processes in heavily petroleum contaminated soils. Six higher plant species (Secale cereale L., Lactuca sativa L., Zea mays L., Lepidium sativum L., Triticum vulgare L., Brassica oleracea L.) were used for bioassay tests based on seed germination and root elongation. The ecotoxicological analyses were made in DMSO/H2O and DCM/DMSO soil extracts. Soils were tested from two biopiles at the Czechowice oil refinery, Poland, that have been subjected to different bioremediation applications. In biopile 1 the active or engineered bioremediation process lasted four years, while biopile 2 was treated passively or non-engineered for eight months. The test species demonstrated varying sensitivity to soils from both biopiles. The effects on test organisms exposed to biopile 2 soils were several times higher compared to those in biopile 1 soils, which correlated with the soil contaminants concentration. Soil hydrocarbon concentrations indeed decreased an average of 81% in biopile 1, whereas in biopile 2 TPH/TPOC concentrations only decreased by 30% after eight months of bioremediation. The bioassays were presented to be sensitive indicators of soil quality and can be used to evaluate the quality of bioremediated soil. The study encourages the need to combine the bioassays with chemical monitoring for evaluation of the bioremediation effectiveness and assessing of the contaminated/remediated soils.     

Effect of hydrocarbon pollution on the microbial properties of a sandy and a clay soil

The aim of this work was to ascertain the effects of different types of hydrocarbon pollution on soil microbial properties and the influence of a soil's characteristics on these effects. For this, toxicity bioassays and microbiological and biochemical parameters were studied in two soils (one sandy and one clayey) contaminated at a loading rate of 5% and 10% with three types of hydrocarbon (diesel oil, gasoline and crude petroleum) differing in their volatilisation potential and toxic substance content. Soils were maintained under controlled conditions (50-70% water holding capacity, and room temperature) for six months and several microbiological and toxicity parameters were monitored 1, 60, 120 and 180 days after contamination. The toxic effects of hydrocarbon contamination were greater in the sandy soil. Hydrocarbons inhibited microbial biomass, the greatest negative effect being observed in the gasoline-polluted sandy soil. In both soils crude petroleum and diesel oil contamination increased microbial respiration, while gasoline had little effect on this parameter, especially in the sandy soil. In general, gasoline had the highest inhibitory effect on the hydrolase activities involved in N, P or C cycles in both soils. All contaminants inhibited hydrolase activities in the sandy soil, while in the clayey soil diesel oil stimulated enzyme activity, particularly at the higher concentration. In both soils, a phytotoxic effect on barley and ryegrass seed germination was observed in the contaminated soils, particularly in those contaminated with diesel or petroleum.     

Soil contamination by crude oil: impact on the mycorrhizosphere and on the revegetation potential of forest trees

In vitro and greenhouse biotests were carried out to study the effects of various concentrations of crude oil on the mycorrhizosphere and the ability of ectomycorrhizal fungi to colonise Norway spruce and poplar seedlings grown on contaminated soil. Ectomycorrhizal fungi grown in pure cultures showed a variety of reactions to crude oil, ranging from growth stimulation to total inhibition of growth, depending on the species of fungi. Germination of poplar and spruce seeds was not significantly affected. The growth of spruce seedlings was not affected by crude oil, whereas that of poplar seedlings was significantly reduced at high concentrations. None of the concentrations had any effect on the degree of ectomycorrhizal and endomycorrhizal colonisation of poplar. With spruce, however, the ectomycorrhizal fungi showed species-specific reactions to increasing concentrations, in accordance with the results of the pure culture test. The length of time between soil contamination and seeding affects both seedling growth and the mycorrhizal infection potential of the soil. The results confirm the importance of mycorrhizal fungi in the bioremediation of soils contaminated by crude oil.     

Air pollution modelling of the Calabrian pine forests withered by the Yatagan thermal power plant, Turkey

The emissions from the chimney of the Yatagan thermal power plant have caused a 3050-hectare forest to wither. Such air pollutants as sulfur dioxide, nitrogen oxides and fly ash are carried towards the Bencikdagi Calabrian pine forests on the prevailing north-east winds. To study these effects, the Q-Basic computer program developed for the Gaussian puff model was used with meteorological data provided by the Yatagan Meteorological Office and the pollutant loads formed by the thermal power plant. In the grid system on the map showing the surroundings of the thermal power plant, the concentrations of the pollutants in each grid square were determined. The Yatagan plant has three units containing 9000 mg/m3 sulfur dioxide, 1800 mg/m3 nitrogen oxides and 680 mg/m3 fly ash in its chimneys. The minimum pollutant emissions were found to be 100 μg/m3 SO₂, 20 μg/m3 NO_x2, and 77 μg/m3 NO_x     

Enhancing bioremediation of diesel oil and gasoline in soil amended with an agroindustry sludge

This paper presents a study of the bioremediation of diesel oil and gasoline by a series of controlled laboratory tests. Sludge from an agroindustry was used to enhance bioremediation of both gasoline and diesel oil mixed with a soil mass to compare its efficiency with that of a mineral fertilizer. Effects of soil microbiology and soil mixtures were investigated by means of evolution of CO2, microorganism populations at 90 days, pH at 65 and 95 days, mineral nitrogen, and gas chromatographic analysis of the benzene, toluene, methyl tertiary butyl ether, C8, and C9+total aromatics at the end of the experiments. Treatments containing sludge showed better soil conditions after 170 days of treatment (inorganic nitrogen and microbiota activity) compared with gasoline and diesel oil without amendments. Samples had no detectable traces of the measured hydrocarbons at 170 days of treatment.     

Influence of sub-lethal crude oil concentration on growth,water relations and photosynthetic capacity of maize (Zea mays L.) plants

Environmental Science and Pollution Research - Maize tolerance potential to oil pollution was assessed by growing Zea mays in soil contaminated with varying levels of crude oil (0, 2.5 and...     

Effects of different ratios of pig manure to fungus residue on physicochemical parameters during composting

This study examined physicochemical parameters to assess their effectiveness as stability and maturity indicators during the process of composting pig manure and fungus residue at different ratios. The results showed that composting mixtures with all ratios of pig manure to fungus residue maintained a temperature exceeding 50 °C for more than 10 days during composting and met the requirement for pathogen destruction. The treatment containing mainly pig manure showed higher nitrogen loss and a shorter thermophilic phase and maturity time than the treatment containing mainly fungus residue. The germination index (GI) values indicated that compost maturity was achieved in the final compost with initial ratios of pig manure to fungus residue of 9:1–7:3 (GIs of 101.4%, 91.2%, and 81.3%); the ratio of 6:4 did not reach compost maturity (GI of 63.8%) and had an inhibitory effect on seed germination. The results of this study suggest that a ratio of pig manure to fungus residue of approximately 8:2 can be considered suitable for the efficient and quality composting of pig manure and fungus residue.

Implications: Co-composting of pig manure and edible fungi residue with appropriate proportion can effectively reduce the risk of environmental pollution caused by agricultural wastes, as well as achieve a safer and high-quality organic fertilizer, which can be used to improve physical and chemical properties of the soil, increase crop yields, and promote agricultural sustainable development. Therefore, technique of co-composting of pig manure and edible fungi residue has a wide prospect of application in practical production all over the world.     

Assessing the hydrocarbon degrading potential of indigenous bacteria isolated from crude oil tank bottom sludge and hydrocarbon-contaminated soil of Azzawiya oil refinery,Libya

The disposal of hazardous crude oil tank bottom sludge (COTBS) represents a significant waste management burden for South Mediterranean countries. Currently, the application of biological systems (bioremediation) for the treatment of COTBS is not widely practiced in these countries. Therefore, this study aims to develop the potential for bioremediation in this region through assessment of the abilities of indigenous hydrocarbonoclastic microorganisms from Libyan Hamada COTBS for the biotreatment of Libyan COTBS-contaminated environments. Bacteria were isolated from COTBS, COTBS-contaminated soil, treated COTBS-contaminated soil, and uncontaminated soil using Bushnell Hass medium amended with Hamada crude oil (1 %) as the main carbon source. Overall, 49 bacterial phenotypes were detected, and their individual abilities to degrade Hamada crude and selected COBTS fractions (naphthalene, phenanthrene, eicosane, octadecane and hexane) were evaluated using MT2 Biolog plates. Analyses using average well colour development showed that ~90 % of bacterial isolates were capable of utilizing representative aromatic fractions compared to 51 % utilization of representative aliphatics. Interestingly, more hydrocarbonoclastic isolates were obtained from treated contaminated soils (42.9 %) than from COTBS (26.5 %) or COTBS-contaminated (30.6 %) and control (0 %) soils. Hierarchical cluster analysis (HCA) separated the isolates into two clusters with microorganisms in cluster 2 being 1.7- to 5-fold better at hydrocarbon degradation than those in cluster 1. Cluster 2 isolates belonged to the putative hydrocarbon-degrading genera; Pseudomonas, Bacillus, Arthrobacter and Brevundimonas with 57 % of these isolates being obtained from treated COTBS-contaminated soil. Overall, this study demonstrates that the potential for PAH degradation exists for the bioremediation of Hamada COTBS-contaminated environments in Libya. This represents the first report on the isolation of hydrocarbonoclastic bacteria from Libyan COTBS and COTBS-contaminated soil.     

利用生物柴油去除滨海构筑物上原油的模拟研究简

溢油向岸滩漂移会造成海岸带人工构筑物的严重污染。采用混凝土片模拟滨海构筑物,研究投加生物柴油及营养对石油污染物的去除效果。结果表明,施加生物柴油可以促进滨海构筑物上的石油进入水中,生物柴油施加量越大,构筑物上残余的石油量越小;同时施加生物柴油和营养能够促进海水中降解石油微生物的增殖和石油的降解;营养和微生物条件一致的情况下,投加2 mL和5 mL生物柴油的系统中石油的总去除率分别为37.5%和32.7%,表明生物柴油的投加量有一个适宜值。研究结果可为生物柴油-营养联合修复石油污染海岸带提供数据支持。     

Electrokinetic enhancement of phenanthrene biodegradation in creosote-polluted clay soil

Given the difficulties caused by low-permeable soils in bioremediation, a new electrokinetic technology is proposed, based on laboratory results with phenanthrene, to afford bioremediation of polycyclic aromatic hydrocarbons (PAH) in clay soils. Microbial activity in a clay soil historically polluted with creosote was promoted using a specially designed electrokinetic cell with a permanent anode-to-cathode flow and controlled pH. The rates of phenanthrene losses during treatment were tenfold higher in soil treated with an electric field than in the control cells without current or microbial activity. Results from experiments with Tenax-assisted desorption and mineralization of 14C-labeled phenanthrene indicated that phenanthrene biodegradation was limited by mass-transfer of the chemical. We suggest that the enhancement effect of the applied electric field on phenanthrene biodegradation resulted from mobilization of the PAH and nutrients dissolved in the soil fluids.     

Rhizoremediation of oil-contaminated sites: a perspective on the Gulf War environmental catastrophe on the State of Kuwait

The Gulf War brought about to the State of Kuwait some of the worst environmental pollution as a result of oil spill. Since 1995, research programs have been initiated to avoid further damage to the Kuwaiti desert and marine environment and to restore and rehabilitate the polluted land, water, and air ecosystems. During the following 15 years, different bioremediation methods both on laboratory and small field scales were tested and evaluated. The findings of these studies were implemented to establish a bio-park in which ornamental shrubs and trees were grown in bioremediated soil. This review will focus on Kuwait’s experience in rhizoremediation and its positive impacts on oil-contaminated sites.     

Effects of spent lubricating oil on the growth of Capsicum annum L. and Lycopersicon esculentum Miller

Susceptibility of the hot pepper (Capsicum annum L.) and the tomato (Lycopersicon esculentum Miller) to spent lubricating oil was investigated in soils supplemented with concentrations of oil ranging from 1-5% w/w. Treatment of soil with 4 and 5% spent oil consistently inhibited germination of both crops. Eighty-four days after sowing, the mean height and leaf area of C. annum in soil treated with 3% spent oil gave the lowest values. These were 3.14 +/- 0.6 cm and 0.24 +/- 0.01 cm(2) compared with respective values of 19.2 +/- 0.2 cm and 2.9 +/- 0.3 cm(2) for control plants. Premature death was observed in the case of L. esculentum plants treated with all tested strengths of spent lubricating oil.     

Biodegradation of aliphatic and aromatic hydrocarbons by natural soil microflora and pure cultures of imperfect and lignolitic fungi

The biodegradation of aliphatic and aromatic hydrocarbons by natural soil microflora and seven fungi species, including imperfect strains and higher level lignolitic species, is compared in a 90-day laboratory experiment using a natural, not-fertilized soil contaminated with 10% crude oil. The natural microbial soil assemblage isolated from an urban forest area was unable to significantly degrade crude oil, whereas pure fungi cultures effectively reduced the residues by 26-35% in 90 days. Normal alkanes were almost completely degraded in the first 15 days, whereas aromatic compounds (phenanthrene and methylphenanthrenes) exhibited slower kinetics. Aspergillus terreus and Fusarium solani, isolated from oil-polluted areas, produced the more efficient attack of aliphatic and aromatic hydrocarbons, respectively. Overall, imperfect fungi isolated from polluted soils showed a somewhat higher efficiency, but the performance of unadapted, indigenous, lignolitic fungi was comparable, and all three species, Pleurotus ostreatus, Trametes villosus and Coriolopsis rigida, effectively degraded aliphatic and aromatic components. The simultaneous, multivariate analysis of 22 parameters allowed the elucidation of a clear reactivity trend of the oil components during biodegradation: lower molecular weight n-alkanes > phenanthrene > 3-2-methylphenanthrenes > intermediate chain length n-alkanes > longer chain length n-alkanes > isoprenoids approximately 9-1-methylphenanthrenes. Irrespective of the individual degrading capacities, all fungi species tested seem to follow this decomposition sequence.     

Effects of nutrient and temperature on degradation of petroleum hydrocarbons in contaminated sub-Antarctic soil

Mesocosm studies using sub-Antarctic soil artificially contaminated with diesel or crude oil were conducted in Kerguelen Archipelago (49 degrees 21' S, 70 degrees 13' E) in an attempt to evaluate the potential of a bioremediation approach in high latitude environments. All mesocosms were sampled on a regular basis over six months period. Soils responded positively to temperature increase from 4 degrees C to 20 degrees C, and to the addition of a commercial oleophilic fertilizer containing N and P. Both factors increased the hydrocarbon-degrading microbial abundance and total petroleum hydrocarbons (TPH) degradation. In general, alkanes were faster degraded than polyaromatic hydrocarbons (PAHs). After 180 days, total alkane losses of both oils reached 77-95% whereas total PAHs never exceeded 80% with optimal conditions at 10 degrees C and fertilizer added. Detailed analysis of naphthalenes, dibenzothiophenes, phenanthrenes, and pyrenes showed a clear decrease of their degradation rate as a function of the size of the PAH molecules. During the experiment there was only a slight decrease in the toxicity, whereas the concentration of TPH decreased significantly during the same time. The most significant reduction in toxicity occurred at 4 degrees C. Therefore, bioremediation of hydrocarbon-contaminated sub-Antarctic soil appears to be feasible, and various engineering strategies, such as heating or amending the soil can accelerate hydrocarbon degradation. However, the residual toxicity of contaminated soil remained drastically high before the desired cleanup is complete and it can represent a limiting factor in the bioremediation of sub-Antarctic soil.     

Rape (Brassica chinensis L.) seed germination, seedling growth, and physiology in soil polluted with di-n-butyl phthalate and bis(2-ethylhexyl) phthalate

Phthalic acid esters (PAEs) pollution in agricultural soils caused by widely employed plastic products is becoming more and more widespread in China. PAEs polluted soil can lead to phytotoxicity in higher plants and potential health risks to human being. We evaluated the individual toxicity of di-n-butyl phthalate (DnBP) and bis(2-ethylhexyl) phthalate (DEHP), two representative PAEs, to sown rape (Brassica chinensis L.) seeds within 72 h (as germination stage) and seedlings after germination for 14 days by monitoring responses and trends of different biological parameters. No significant effects of six concentrations of PAE ranging from 0 (not treated/NT) to 500 mg?kg^?1 on germination rate in soil were observed. However, root length, shoot length, and biomass (fresh weight) were inhibited by both pollutants (except root length and biomass under DEHP). Stimulatory effects of both target pollutants on malondialdehyde (MDA) content, superoxide dismutase (SODase) activity, ascorbate peroxidase (APXase) content, and polyphenoloxidase (PPOase) activity in shoots and roots (SODase activity in shoots excluded) were in the same trend with the promotion of proline (Pro) but differed with acetylcholinesterase activity (except in shoots under DnBP) for analyzed samples treated for 72 h and 14 days. Responses of representative storage compounds free amino acids (FAA) and total soluble sugar (TSS) under both PAEs were raised. Sensitivity of APXase and Pro in roots demonstrates their possibility in estimation of PAE phytotoxicity and the higher toxicity of DnBP, which has also been approved by the morphological photos of seedlings at day 14. Higher sensitivity of the roots was also observed. The recommended soil allowable concentration is 5 mg DnBP?kg^?1 soil for the development of rape. We still need to know the phytotoxicity of DEHP at whole seedling stage for both the growing and development; on the other hand, soil criteria for PAE compounds are urgently required in China.